β lactamase Search Results


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  • 95
    Millipore β lactamase extract
    Analysis of TEM-67 purification by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with Coomassie blue staining. Lane 1, crude <t>β-lactamase</t> extract; lane 2, protein extract after first Q-Sepharose column; lane 3, purified protein. Molecular size markers were run in lane M, and their sizes are indicated on the right.
    β Lactamase Extract, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 56 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/β lactamase extract/product/Millipore
    Average 95 stars, based on 56 article reviews
    Price from $9.99 to $1999.99
    β lactamase extract - by Bioz Stars, 2020-08
    95/100 stars
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    92
    Millipore β lactamase
    Rubcn -/- DCs exhibit increased phagosome-to-cytosol escape and proteasome-mediated generation of peptides. Bone marrow-derived dendritic cells (DCs) were generated from Rubcn +/+ (black) and Rubcn -/- (red) mice in vitro with FLT3-L for 7 days. ( A-B ) DCs were loaded with 1 µm CCF4, then co-cultured with apoptotic B16-OVA in the absence or presence of <t>β-lactamase</t> (2 mg/ml) for 90 or 180 minutes at 4°C or 37°C. Uncleaved CCF4 was measured by flow cytometry at an emission of 535 nm, and cleaved CCF4 was measured at an emission of 450 nm. Ratios of 450 nm : 535 nm was calculated by dividing the 450 nm MFI by the 535 nm MFI from a single sample. ( C ) DCs were pre-treated with vehicle or chloroquine (CQ) at 1 or 10 µM for 2 hours and then co-cultured with apoptotic B16-OVA cells (5 apoptotic cells: 1 DC). Eighteen hours later, DCs were harvested for flow cytometry analysis of H2-K b -OVA 257-264 expression. ( D ) DCs were pre-treated with vehicle or MG-132 at 1 or 10 µM for 2 hours and then co-cultured in fresh media with apoptotic B16-OVA cells (5 apoptotic cells: 1 DC). Eighteen hours later, DCs were harvested for flow cytometry analysis of H2-K b -OVA 257-264 expression. ( E ) DCs were pre-treated with vehicle or Brefeldin A at 3 µg/ml for 2 hours and then co-cultured in fresh media with apoptotic B16-OVA cells (5 apoptotic cells: 1 DC). Eighteen hours later, DCs were harvested for flow cytometry analysis of H2-K b -OVA 257-264 expression. Data are expressed as mean ± SEM. No less than two independent experiments were performed, with 3-5 replicates per condition. Significance was calculated using 2-way ANOVA (*p
    β Lactamase, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 300 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/β lactamase/product/Millipore
    Average 92 stars, based on 300 article reviews
    Price from $9.99 to $1999.99
    β lactamase - by Bioz Stars, 2020-08
    92/100 stars
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    99
    Millipore β lactamase type iv
    The inhibitory activity of quercetin against <t>β-lactamase</t> in hydrolyzing benzylpenicillin. β-lactamase used from E. cloacae ; Con = control (no testing agent); Que (50) = Quercetin 50 μg/mL. The graph shows the remaining benzylpenicillin at the same time. Means sharing the same superscript are not significantly different from each other (Scheffe’s test, p
    β Lactamase Type Iv, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/β lactamase type iv/product/Millipore
    Average 99 stars, based on 15 article reviews
    Price from $9.99 to $1999.99
    β lactamase type iv - by Bioz Stars, 2020-08
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    Image Search Results


    Analysis of TEM-67 purification by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with Coomassie blue staining. Lane 1, crude β-lactamase extract; lane 2, protein extract after first Q-Sepharose column; lane 3, purified protein. Molecular size markers were run in lane M, and their sizes are indicated on the right.

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Integration of a Transposon Tn1-Encoded Inhibitor-Resistant ?-Lactamase Gene, blaTEM-67 from Proteus mirabilis, into the Escherichia coli Chromosome

    doi: 10.1128/AAC.47.1.19-26.2003

    Figure Lengend Snippet: Analysis of TEM-67 purification by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with Coomassie blue staining. Lane 1, crude β-lactamase extract; lane 2, protein extract after first Q-Sepharose column; lane 3, purified protein. Molecular size markers were run in lane M, and their sizes are indicated on the right.

    Article Snippet: The β-lactamase extract was filtered through a 0.45-μm-pore-size filter (Millipore, Saint-Quentin-en-Yvelines, France) and ultrafiltrated with a Vivaspin 100,000 column (Sartorius, Gottingen, Germany) prior to being loaded onto a preequilibrated Q-Sepharose column (Amersham Pharmacia Biotech) in 20 mM Tris HCl buffer (pH 7.5).

    Techniques: Transmission Electron Microscopy, Purification, Polyacrylamide Gel Electrophoresis, Staining

    Purified β-lactamase proteins. The six β-lactamase proteins were purified to > 90% homogeneity, as determined by SDS-PAGE. The values on the molecular weight ladder (MWL) are expressed in units of kilodaltons.

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Binding Properties of a Peptide Derived from ?-Lactamase Inhibitory Protein

    doi: 10.1128/AAC.45.12.3279-3286.2001

    Figure Lengend Snippet: Purified β-lactamase proteins. The six β-lactamase proteins were purified to > 90% homogeneity, as determined by SDS-PAGE. The values on the molecular weight ladder (MWL) are expressed in units of kilodaltons.

    Article Snippet: The β-lactamase gene was induced by adding isopropyl-β- d -thiogalactopyranoside (IPTG) to a final concentration of 0.5 mM and further incubation at 25°C for 4 h. Following induction the cells were pelleted and the supernatant containing the secreted, soluble β-lactamase was concentrated to 100 ml with an Amicon Centriprep-10 concentrator (Millipore Corp.).

    Techniques: Purification, SDS Page, Molecular Weight

    ). Closed squares, protein kinase C peptide; open squares, cyclic BP46-51 peptide; closed circles, reduced BP46-51 peptide; open circles, BP41-50 peptide. Each datum point is the average for two independent experiments. (B) Inhibition assay of TEM-1 β-lactamase by wild-type BLIP. A K i of 0.23 nM was determined by using a nonlinear regression fit as described above.

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Binding Properties of a Peptide Derived from ?-Lactamase Inhibitory Protein

    doi: 10.1128/AAC.45.12.3279-3286.2001

    Figure Lengend Snippet: ). Closed squares, protein kinase C peptide; open squares, cyclic BP46-51 peptide; closed circles, reduced BP46-51 peptide; open circles, BP41-50 peptide. Each datum point is the average for two independent experiments. (B) Inhibition assay of TEM-1 β-lactamase by wild-type BLIP. A K i of 0.23 nM was determined by using a nonlinear regression fit as described above.

    Article Snippet: The β-lactamase gene was induced by adding isopropyl-β- d -thiogalactopyranoside (IPTG) to a final concentration of 0.5 mM and further incubation at 25°C for 4 h. Following induction the cells were pelleted and the supernatant containing the secreted, soluble β-lactamase was concentrated to 100 ml with an Amicon Centriprep-10 concentrator (Millipore Corp.).

    Techniques: Inhibition, Transmission Electron Microscopy

    Representation of the secondary structures surrounding the amino acid at position 148. Left panel, Native AmpC β-lactamase of C. freundii GN346 (PDB 1RGY ); Right panel, AmpC variant of the AmpC β-lactamase of C. freundii GN346 presenting

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Hydrolysis Spectrum Extension of CMY-2-Like ?-Lactamases Resulting from Structural Alteration in the Y-X-N Loop

    doi: 10.1128/AAC.05630-11

    Figure Lengend Snippet: Representation of the secondary structures surrounding the amino acid at position 148. Left panel, Native AmpC β-lactamase of C. freundii GN346 (PDB 1RGY ); Right panel, AmpC variant of the AmpC β-lactamase of C. freundii GN346 presenting

    Article Snippet: E. coli TOP10 recombinant clones were grown overnight at 37°C in 4 liters of Trypticase soy (TS) broth containing amoxicillin (100 μg/ml) and kanamycin (30 μg/ml), resuspended in 40 ml of 100 mM phosphate buffer (pH 7), disrupted by sonication, and centrifuged at 20,000 × g for 1 h at 4°C, as described previously ( ). β-Lactamase extracts were filtered through a 0.45-mm-pore-size filter (Millipore, Saint-Quentin-en-Yvelines, France), dialyzed overnight at 4°C against 20 mM Tris (pH 7.5), and loaded on a preequilibrated Q-Sepharose column (Amersham Pharmacia Biotech).

    Techniques: Variant Assay

    Optimization of Shigella T3SS injection reporter. ( A ) Immunoblotting with anti–β-lactamase antibody of lysates ( Top ) and supernatants (sups.; Bottom ) of ipaD strains expressing TEM-1 chimeric proteins. The expected molecular weights of chimeric proteins were as follows (in kDa): IpgD-bla (89), OspC2-bla (85), IpaJ-bla (58), OspD1-bla (54), OspD1sh-bla (sh-bla) (38), OspD1sh-M31L-bla (sh-M31L-bla) (38), and OspD1ctrl-bla (ctrl-bla) (35). ( B ) Measure of β-lactamase activity in the ipaD supernatants after incubation with nitrocefin. Data are from three independent experiments. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001 (Student’s t test compared with OspD1ctrl-bla condition, unless depicted otherwise). ( C ) Measure of β-lactamase activity in the supernatants of ipaD expressing OspD1sh-M31L fused to TEM variants after incubation with CCF2-FA. Data are from four independent experiments. * P ≤ 0.05 (Student’s t test compared with TEM1 condition). ( D ) Flow cytometry analysis of CCF2-AM–loaded Jurkat T lymphocytes infected for 1 h at an MOI of 50 with WT Shigella expressing the optimized T3SS injection reporter with or without DsRed. Data are from five independent experiments.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Injection of T3SS effectors not resulting in invasion is the main targeting mechanism of Shigella toward human lymphocytes

    doi: 10.1073/pnas.1707098114

    Figure Lengend Snippet: Optimization of Shigella T3SS injection reporter. ( A ) Immunoblotting with anti–β-lactamase antibody of lysates ( Top ) and supernatants (sups.; Bottom ) of ipaD strains expressing TEM-1 chimeric proteins. The expected molecular weights of chimeric proteins were as follows (in kDa): IpgD-bla (89), OspC2-bla (85), IpaJ-bla (58), OspD1-bla (54), OspD1sh-bla (sh-bla) (38), OspD1sh-M31L-bla (sh-M31L-bla) (38), and OspD1ctrl-bla (ctrl-bla) (35). ( B ) Measure of β-lactamase activity in the ipaD supernatants after incubation with nitrocefin. Data are from three independent experiments. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001 (Student’s t test compared with OspD1ctrl-bla condition, unless depicted otherwise). ( C ) Measure of β-lactamase activity in the supernatants of ipaD expressing OspD1sh-M31L fused to TEM variants after incubation with CCF2-FA. Data are from four independent experiments. * P ≤ 0.05 (Student’s t test compared with TEM1 condition). ( D ) Flow cytometry analysis of CCF2-AM–loaded Jurkat T lymphocytes infected for 1 h at an MOI of 50 with WT Shigella expressing the optimized T3SS injection reporter with or without DsRed. Data are from five independent experiments.

    Article Snippet: Subcultures were spun at 9,000 × g for 1 min, and 100 μL of supernatants at different dilutions were incubated with β-lactamase substrates: 20 μL of nitrocefin (500 μg/mL stock; EMD Millipore) or 20 μL of CCF2-FA (25 μM stock; Thermo Fisher Scientific).

    Techniques: Injection, Expressing, Transmission Electron Microscopy, Activity Assay, Incubation, Flow Cytometry, Cytometry, Infection

    Proposed mechanism of the retroaldolic reaction leading to elimination of C 6 hydroxethyl substituent from the β-lactamase: carbapenem acyl-enzyme. Glu272, supported by Lys315, may serve as the base to deprotonate the alcoholic function β–hydroxy

    Journal: Biochemistry

    Article Title: Inhibition of the Class C ?-Lactamase from Acinetobacter spp: Insights into Effective Inhibitor Design †

    doi: 10.1021/bi9015988

    Figure Lengend Snippet: Proposed mechanism of the retroaldolic reaction leading to elimination of C 6 hydroxethyl substituent from the β-lactamase: carbapenem acyl-enzyme. Glu272, supported by Lys315, may serve as the base to deprotonate the alcoholic function β–hydroxy

    Article Snippet: For large-scale protein expression and β-lactamase characterization, the bla ADC gene (specifically, bla ADC-7 ) was cloned into pET24a (+) vector (kanamycin resistant, Novagen, Madison, WI) following a previously published method ( ).

    Techniques:

    Deconvoluted mass spectra of: (A) ADC β–lactamase alone; (B) ADC after 15 min incubation with compounds 2 and 5 ; and (C) ADC β-lactamase after 15 min incubation with imipenem, ertapenem, doripenem, and meropenem. The peak in each

    Journal: Biochemistry

    Article Title: Inhibition of the Class C ?-Lactamase from Acinetobacter spp: Insights into Effective Inhibitor Design †

    doi: 10.1021/bi9015988

    Figure Lengend Snippet: Deconvoluted mass spectra of: (A) ADC β–lactamase alone; (B) ADC after 15 min incubation with compounds 2 and 5 ; and (C) ADC β-lactamase after 15 min incubation with imipenem, ertapenem, doripenem, and meropenem. The peak in each

    Article Snippet: For large-scale protein expression and β-lactamase characterization, the bla ADC gene (specifically, bla ADC-7 ) was cloned into pET24a (+) vector (kanamycin resistant, Novagen, Madison, WI) following a previously published method ( ).

    Techniques: Incubation

    Schemes illustrating the interactions of a serine β-lactamase with: (A) the β-lactam cephalosporin ceftazidime; (B) the boronic acid ceftazidime analog, compound 2 ; and (C) the carbapenem imipenem.

    Journal: Biochemistry

    Article Title: Inhibition of the Class C ?-Lactamase from Acinetobacter spp: Insights into Effective Inhibitor Design †

    doi: 10.1021/bi9015988

    Figure Lengend Snippet: Schemes illustrating the interactions of a serine β-lactamase with: (A) the β-lactam cephalosporin ceftazidime; (B) the boronic acid ceftazidime analog, compound 2 ; and (C) the carbapenem imipenem.

    Article Snippet: For large-scale protein expression and β-lactamase characterization, the bla ADC gene (specifically, bla ADC-7 ) was cloned into pET24a (+) vector (kanamycin resistant, Novagen, Madison, WI) following a previously published method ( ).

    Techniques:

    BLAST delivered β-lactamase enzyme is functional inside NHDFs

    Journal: Nature methods

    Article Title: Massively Parallel Delivery of Large-Sized Cargo into Mammalian Cells with Light Pulses

    doi: 10.1038/nmeth.3357

    Figure Lengend Snippet: BLAST delivered β-lactamase enzyme is functional inside NHDFs

    Article Snippet: Mouse anti-α tubulin with Alexa Fluor 488 (abcam) was prepared to 100 µg/ml in PBS. β-lactamase enzyme from Enterobacter cloacae (Sigma) was dissolved in PBS at 50 units/ml.

    Techniques: Functional Assay

    Relative β‐lactamase activities of wall‐bound fusions on the surface of intact cells and in the cell wall fraction. Cell wall fraction is defined as the supernatant fraction generated after centrifugal removal of protoplasts from the intact cell samples that have been treated with lysozyme. β‐Lactamase activity was determined in the presence of BSA. The activity from the cell wall fraction of E48B was set as 100%. Three independent experiments were carried. CWBM LytE : cell wall‐binding module from LytE. Bla: secreted TEM‐β‐lactamase. This is the mature form of β‐lactamase in the culture supernatant of WB800[pWB980‐Bla]. Secretion of this protein is directed by the B. subtilis SacB signal peptide.

    Journal: Microbial Biotechnology

    Article Title: Development of a LytE-based high-density surface display system in Bacillus subtilis

    doi: 10.1111/j.1751-7915.2007.00017.x

    Figure Lengend Snippet: Relative β‐lactamase activities of wall‐bound fusions on the surface of intact cells and in the cell wall fraction. Cell wall fraction is defined as the supernatant fraction generated after centrifugal removal of protoplasts from the intact cell samples that have been treated with lysozyme. β‐Lactamase activity was determined in the presence of BSA. The activity from the cell wall fraction of E48B was set as 100%. Three independent experiments were carried. CWBM LytE : cell wall‐binding module from LytE. Bla: secreted TEM‐β‐lactamase. This is the mature form of β‐lactamase in the culture supernatant of WB800[pWB980‐Bla]. Secretion of this protein is directed by the B. subtilis SacB signal peptide.

    Article Snippet: The pooled β‐lactamase fractions were concentrated using the Ultrafree column (Millipore) prior to applying to a BioPrep SE100/17 gel filtration column (BioRad) equilibrated with the SPSC‐1 buffer (20 mM sodium phosphate, pH 8.0, 0.15 M sodium chloride).

    Techniques: Generated, Activity Assay, Binding Assay, Transmission Electron Microscopy

    Construction and properties of CWBM LytE and its fusions. A. Constructs of CWBM LytE (black), β‐lactamase (Bla, white) and their fusions. Linker regions are highlighted in grey. L5, L32, L48 and L55 indicate the presence of 5, 32, 48 and 55 amino acids in the linker regions respectively. Constructs 3 (B5E, B for β‐lactamase and E for CWBM LytE ), 4 (B32E) and 5 (B55E) have the β‐lactamase at the N‐terminal end. They are the members of the BE series. Construct 6 (E48B) has the β‐lactamase at the C‐terminal end. M M indicates molecular mass. B. Coomassie blue‐stained SDS‐polyacrylamide gel (12%) profile showing the apparent molecular masses and purities of the purified CWBM LytE , β‐lactamase and their fusions. M represents the lane loaded with molecular mass markers.

    Journal: Microbial Biotechnology

    Article Title: Development of a LytE-based high-density surface display system in Bacillus subtilis

    doi: 10.1111/j.1751-7915.2007.00017.x

    Figure Lengend Snippet: Construction and properties of CWBM LytE and its fusions. A. Constructs of CWBM LytE (black), β‐lactamase (Bla, white) and their fusions. Linker regions are highlighted in grey. L5, L32, L48 and L55 indicate the presence of 5, 32, 48 and 55 amino acids in the linker regions respectively. Constructs 3 (B5E, B for β‐lactamase and E for CWBM LytE ), 4 (B32E) and 5 (B55E) have the β‐lactamase at the N‐terminal end. They are the members of the BE series. Construct 6 (E48B) has the β‐lactamase at the C‐terminal end. M M indicates molecular mass. B. Coomassie blue‐stained SDS‐polyacrylamide gel (12%) profile showing the apparent molecular masses and purities of the purified CWBM LytE , β‐lactamase and their fusions. M represents the lane loaded with molecular mass markers.

    Article Snippet: The pooled β‐lactamase fractions were concentrated using the Ultrafree column (Millipore) prior to applying to a BioPrep SE100/17 gel filtration column (BioRad) equilibrated with the SPSC‐1 buffer (20 mM sodium phosphate, pH 8.0, 0.15 M sodium chloride).

    Techniques: Construct, Staining, Purification

    Immobilization of enzymes on the surface of lactococcal GEM particles. (a) Relative α-amylase activities on GEM particles incubated with culture medium containing soluble AmyL, PA (PA3), or α-PA. (b) Relative α-amylase and β-lactamase

    Journal: Applied and Environmental Microbiology

    Article Title: Novel Surface Display System for Proteins on Non-Genetically Modified Gram-Positive Bacteria

    doi: 10.1128/AEM.72.1.880-889.2006

    Figure Lengend Snippet: Immobilization of enzymes on the surface of lactococcal GEM particles. (a) Relative α-amylase activities on GEM particles incubated with culture medium containing soluble AmyL, PA (PA3), or α-PA. (b) Relative α-amylase and β-lactamase

    Article Snippet: After 60 min, GEM particles and insoluble amylose azure were spun down, and the absorbance at 595 nm was determined. β-Lactamase activity was measured by adding 40 μl nitrocefin (CalBiochem) to GEM particles loaded with β-PA in 1 ml (final volume) of PBS.

    Techniques: Incubation

    Protein synthesis, miscoding and ETC are impaired by deletions of rbbA and yhjD . (A) Incorporation of [ 35 S]-methionine into total proteins of the wild type and the indicated deletion strains in the absence and presence of sodium azide (500 µM) or CCCP (10 µM). Error bars indicate mean ± SD. (B) Elevated rates of frame-shifting and stop-codon read-through in wild type and in the indicated deletion strains in the absence and presence of sodium azide (500 µM) or CCCP (10 µM). Error bars indicate mean ± SD. The β-gal activity is normalized to the activity of β-lactamase that has been used as an internal control. (C) Immunoblot of the full-length and truncated β-gal is assayed after transforming the reporter plasmid pSG400 carrying a premature UAG stop codon expressed in rbbA-yhjD double mutant (Lane 4) and in the rbbA (Lane 2) and yhjD (Lane 3) single mutants. The assay was also performed with another reporter plasmid 4pCCCU containing +1 frame-shift expressed in rbbA - yhjD double mutant (Lane 8) and in the rbbA (Lane 6) and yhjD (Lane 7) single mutants. The wild type and plasmids pSG400 (Lane 1) and 4pCCCU (Lane 5) served as controls. (D) Growth sensitivity assay of the indicated deletion strains in the presence or absence of antibiotics affecting miscoding, translocation and accuracy of protein synthesis. The minimum inhibitory concentration (MIC) of the antibiotic is shown in parenthesis. The hypomorphic allele of the essential gene fusA and the non-essential mutant tufB served as controls. (E) Serial-dilution assay showing the reduced growth fitness defect of rbbA - yhjD double mutants and their corresponding single mutants on M9 minimal media in the presence of 2% glucose or glycerol. (F) Growth sensitivity of the indicated deletion strains in the presence or absence of piericidin A targeting the NDH (Complex I) in ETC. The MIC of the drug is shown in parenthesis. The nuoL mutant served as control.

    Journal: PLoS ONE

    Article Title: Ribosome-Dependent ATPase Interacts with Conserved Membrane Protein in Escherichia coli to Modulate Protein Synthesis and Oxidative Phosphorylation

    doi: 10.1371/journal.pone.0018510

    Figure Lengend Snippet: Protein synthesis, miscoding and ETC are impaired by deletions of rbbA and yhjD . (A) Incorporation of [ 35 S]-methionine into total proteins of the wild type and the indicated deletion strains in the absence and presence of sodium azide (500 µM) or CCCP (10 µM). Error bars indicate mean ± SD. (B) Elevated rates of frame-shifting and stop-codon read-through in wild type and in the indicated deletion strains in the absence and presence of sodium azide (500 µM) or CCCP (10 µM). Error bars indicate mean ± SD. The β-gal activity is normalized to the activity of β-lactamase that has been used as an internal control. (C) Immunoblot of the full-length and truncated β-gal is assayed after transforming the reporter plasmid pSG400 carrying a premature UAG stop codon expressed in rbbA-yhjD double mutant (Lane 4) and in the rbbA (Lane 2) and yhjD (Lane 3) single mutants. The assay was also performed with another reporter plasmid 4pCCCU containing +1 frame-shift expressed in rbbA - yhjD double mutant (Lane 8) and in the rbbA (Lane 6) and yhjD (Lane 7) single mutants. The wild type and plasmids pSG400 (Lane 1) and 4pCCCU (Lane 5) served as controls. (D) Growth sensitivity assay of the indicated deletion strains in the presence or absence of antibiotics affecting miscoding, translocation and accuracy of protein synthesis. The minimum inhibitory concentration (MIC) of the antibiotic is shown in parenthesis. The hypomorphic allele of the essential gene fusA and the non-essential mutant tufB served as controls. (E) Serial-dilution assay showing the reduced growth fitness defect of rbbA - yhjD double mutants and their corresponding single mutants on M9 minimal media in the presence of 2% glucose or glycerol. (F) Growth sensitivity of the indicated deletion strains in the presence or absence of piericidin A targeting the NDH (Complex I) in ETC. The MIC of the drug is shown in parenthesis. The nuoL mutant served as control.

    Article Snippet: Cell pellets were resuspended in 0.1 M phosphate buffer pH 7.5 and lysed by sonication. β-galactosidase assays were performed using the O-nitrophenyl-α-D-galactopyranoside (ONPG) method and the β-lactamase activities were measured using a CENTA kit (Calbiochem).

    Techniques: Activity Assay, Plasmid Preparation, Mutagenesis, Sensitive Assay, Translocation Assay, Concentration Assay, Serial Dilution Assay

    Dendrogram showing pulsed-field gel electrophoresis (PFGE) analysis and multilocus sequence typing (MLST) results for New Delhi metallo-β-lactamase–producing Klebsiella pneumoniae isolates, United States, April 2009–March 2011. Scale bar indicates % similarity. CA, California; MD, Maryland.

    Journal: Emerging Infectious Diseases

    Article Title: New Delhi Metallo-?-Lactamase-producing Enterobacteriaceae, United States

    doi: 10.3201/eid1906.121515

    Figure Lengend Snippet: Dendrogram showing pulsed-field gel electrophoresis (PFGE) analysis and multilocus sequence typing (MLST) results for New Delhi metallo-β-lactamase–producing Klebsiella pneumoniae isolates, United States, April 2009–March 2011. Scale bar indicates % similarity. CA, California; MD, Maryland.

    Article Snippet: BMD Screening for Metallo-β-Lactamase MICs in the absence and presence of a combination of 0.2 mmol/L EDTA (Sigma-Aldrich, St. Louis, MO, USA) and 0.02 mmol/L 1,10-phenanthroline (Acros Organics, Geel, Belgium) were determined as described ( ) by using screening wells containing IMP at concentrations ranging from 0.25 µg/mL through 1,024 µg/mL.

    Techniques: Pulsed-Field Gel, Electrophoresis, Sequencing

    Xmn I restriction analysis of New Delhi metallo-β-lactamase (NDM)–encoding plasmids, United States, April 2009–March 2011, from transformants (A) and subsequent Southern blot analysis with digoxigenin-labeled bla NDM probe hybridized to a blot of same gel (B). Lane 1, NDM PCR product, positive control; lane 2, NDM-negative plasmid (ATCC-1705); lanes 3 and 14, 1-kb plus marker; lane 4, TF 0S-506; lane 5, TF 1100770; lane 6, TF 1100975; lane 7, TF1100192; lane 8, TF 1000527; lane 9, TF 1101459; lane 10, TF 1101168; lane 11, TF 1100101; lane 12, TF 1001728; lane 13, TF 1000654.

    Journal: Emerging Infectious Diseases

    Article Title: New Delhi Metallo-?-Lactamase-producing Enterobacteriaceae, United States

    doi: 10.3201/eid1906.121515

    Figure Lengend Snippet: Xmn I restriction analysis of New Delhi metallo-β-lactamase (NDM)–encoding plasmids, United States, April 2009–March 2011, from transformants (A) and subsequent Southern blot analysis with digoxigenin-labeled bla NDM probe hybridized to a blot of same gel (B). Lane 1, NDM PCR product, positive control; lane 2, NDM-negative plasmid (ATCC-1705); lanes 3 and 14, 1-kb plus marker; lane 4, TF 0S-506; lane 5, TF 1100770; lane 6, TF 1100975; lane 7, TF1100192; lane 8, TF 1000527; lane 9, TF 1101459; lane 10, TF 1101168; lane 11, TF 1100101; lane 12, TF 1001728; lane 13, TF 1000654.

    Article Snippet: BMD Screening for Metallo-β-Lactamase MICs in the absence and presence of a combination of 0.2 mmol/L EDTA (Sigma-Aldrich, St. Louis, MO, USA) and 0.02 mmol/L 1,10-phenanthroline (Acros Organics, Geel, Belgium) were determined as described ( ) by using screening wells containing IMP at concentrations ranging from 0.25 µg/mL through 1,024 µg/mL.

    Techniques: Southern Blot, Labeling, Polymerase Chain Reaction, Positive Control, Plasmid Preparation, Marker

    Total primary and  β -lactamase-active primary clones in the randomized-loop libraries

    Journal: Journal of molecular recognition : JMR

    Article Title: Developing bifunctional β-lactamase molecules with built-in target-recognizing module for prodrug therapy: identification of Enterobacter Cloacae P99 cephalosporinase loops suitable for randomization and phage-display selection

    doi: 10.1002/jmr.957

    Figure Lengend Snippet: Total primary and β -lactamase-active primary clones in the randomized-loop libraries

    Article Snippet: These samples were normalized for their β -lactamase levels by dilution in PBS solution containing bacterial protease inhibitor cocktail (Sigma Chemical Company).

    Techniques: Clone Assay

    Screening of β -lactamase clones selected from the loop-2 linear library panning for their binding to streptavidin (target) and BSA (control) proteins. The bars represent the mean ± SE of the two experiments done in triplicate. The binding

    Journal: Journal of molecular recognition : JMR

    Article Title: Developing bifunctional β-lactamase molecules with built-in target-recognizing module for prodrug therapy: identification of Enterobacter Cloacae P99 cephalosporinase loops suitable for randomization and phage-display selection

    doi: 10.1002/jmr.957

    Figure Lengend Snippet: Screening of β -lactamase clones selected from the loop-2 linear library panning for their binding to streptavidin (target) and BSA (control) proteins. The bars represent the mean ± SE of the two experiments done in triplicate. The binding

    Article Snippet: These samples were normalized for their β -lactamase levels by dilution in PBS solution containing bacterial protease inhibitor cocktail (Sigma Chemical Company).

    Techniques: Clone Assay, Binding Assay

    Randomization of selected P99 β -lactamase loops for the construction different libraries

    Journal: Journal of molecular recognition : JMR

    Article Title: Developing bifunctional β-lactamase molecules with built-in target-recognizing module for prodrug therapy: identification of Enterobacter Cloacae P99 cephalosporinase loops suitable for randomization and phage-display selection

    doi: 10.1002/jmr.957

    Figure Lengend Snippet: Randomization of selected P99 β -lactamase loops for the construction different libraries

    Article Snippet: These samples were normalized for their β -lactamase levels by dilution in PBS solution containing bacterial protease inhibitor cocktail (Sigma Chemical Company).

    Techniques:

    Randomized loops of  β -lactamase. (A) Schematic representation of different loops (solid black) of the  Enterobacter cloacae ), that were

    Journal: Journal of molecular recognition : JMR

    Article Title: Developing bifunctional β-lactamase molecules with built-in target-recognizing module for prodrug therapy: identification of Enterobacter Cloacae P99 cephalosporinase loops suitable for randomization and phage-display selection

    doi: 10.1002/jmr.957

    Figure Lengend Snippet: Randomized loops of β -lactamase. (A) Schematic representation of different loops (solid black) of the Enterobacter cloacae ), that were

    Article Snippet: These samples were normalized for their β -lactamase levels by dilution in PBS solution containing bacterial protease inhibitor cocktail (Sigma Chemical Company).

    Techniques:

    Affinity of Pet for purified VirK. (A) E. coli BL21 was fractionated as described in Materials and Methods. Cytoplasmic, IM, periplasmic, and OM fractions were analyzed by Western blot (WB) assay using anti-His antibodies and, as markers, antibodies against GroEL and β-lactamase. (B) Bacterial cultures transformed with pVirK-His were lysed 4 h after IPTG induction. VirK-His 6 was purified from cell lysates supernatant by Ni-NTA agarose affinity chromatography in accordance with the manufacturer's instructions (Qiagen). Fractions from different stages of the purification process were resolved by SDS-PAGE and transferred to a nitrocellulose membrane. VirK-His 6 was visualized using a mouse anti-His primary antibody and an HRP-conjugated anti-mouse IgG secondary antibody. The arrow indicates the recombinant VirK protein. (C and D) Purified Pet, purified and denatured Pet, the purified β-domain, or the purified and denatured β-domain was incubated with VirK-His. Parallel samples were incubated with fractions 2 and 3 from BL21 transformed with the empty vector (pRSETA) as a negative control. After an overnight incubation, the complexes were immunoprecipitated with anti-His antibodies. As an additional control, the antibodies were incubated in the absence of either Pet or the β domain. The immunocomplexes (C) and nonimmunoprecipitated proteins (D) were separated by SDS-PAGE and visualized with Coomassie brilliant blue stain. MWM, MW markers.

    Journal: Infection and Immunity

    Article Title: VirK Is a Periplasmic Protein Required for Efficient Secretion of Plasmid-Encoded Toxin from Enteroaggregative Escherichia coli

    doi: 10.1128/IAI.00167-12

    Figure Lengend Snippet: Affinity of Pet for purified VirK. (A) E. coli BL21 was fractionated as described in Materials and Methods. Cytoplasmic, IM, periplasmic, and OM fractions were analyzed by Western blot (WB) assay using anti-His antibodies and, as markers, antibodies against GroEL and β-lactamase. (B) Bacterial cultures transformed with pVirK-His were lysed 4 h after IPTG induction. VirK-His 6 was purified from cell lysates supernatant by Ni-NTA agarose affinity chromatography in accordance with the manufacturer's instructions (Qiagen). Fractions from different stages of the purification process were resolved by SDS-PAGE and transferred to a nitrocellulose membrane. VirK-His 6 was visualized using a mouse anti-His primary antibody and an HRP-conjugated anti-mouse IgG secondary antibody. The arrow indicates the recombinant VirK protein. (C and D) Purified Pet, purified and denatured Pet, the purified β-domain, or the purified and denatured β-domain was incubated with VirK-His. Parallel samples were incubated with fractions 2 and 3 from BL21 transformed with the empty vector (pRSETA) as a negative control. After an overnight incubation, the complexes were immunoprecipitated with anti-His antibodies. As an additional control, the antibodies were incubated in the absence of either Pet or the β domain. The immunocomplexes (C) and nonimmunoprecipitated proteins (D) were separated by SDS-PAGE and visualized with Coomassie brilliant blue stain. MWM, MW markers.

    Article Snippet: The quality of the purified periplasmic fractions was tested by Western blot assay using anti-GroEL (kindly donated by Mario Cancino) and anti-β-lactamase (Chemicon, Temecula, CA) antibodies to detect GroEL (a cytoplasmic protein) and β-lactamase (a periplasmic protein), respectively.

    Techniques: Positron Emission Tomography, Purification, Western Blot, Transformation Assay, Affinity Chromatography, SDS Page, Recombinant, Incubation, Plasmid Preparation, Negative Control, Immunoprecipitation, Staining

    Pet interacts with specific periplasmic proteins from E. coli HB101 during secretion. (A) Bacterial strains grown overnight at 37°C at 150 rpm in 50 ml of LB broth were fractionated as described in Materials and Methods. Cytoplasmic, IM, periplasmic, and OM fractions were analyzed by Western blot (WB) assay using as markers antibodies against GroEL, β-lactamase, and OmpA. (B) The periplasmic fraction of E. coli HB101 was immunoprecipitated with an anti-Pet polyclonal antibody. Experiments were performed with an untransformed laboratory strain that does not express Pet (HB101), a strain transformed with a plasmid encoding wild-type Pet (pCEFN1), a strain transformed with a plasmid encoding the inactive mutant protein PetS260I (pCEFN2), and a strain transformed with a plasmid encoding the truncated mutant Pet protein that lacks the β-barrel translocation unit (pJPN205). An additional negative control involved coimmunoprecipitation of the periplasmic fraction from HB101(pCEFN1) with an isotype IgG control antibody. All immunocomplexes were resolved by SDS-PAGE and visualized by Coomassie blue staining. Arrows indicate the proteins (20, 37 50, and 80 kDa) that specifically interacted with PetS260I; asterisks denote the 50- and 80-kDa proteins that also bound to the truncated mutant Pet protein. MWM, MW markers.

    Journal: Infection and Immunity

    Article Title: VirK Is a Periplasmic Protein Required for Efficient Secretion of Plasmid-Encoded Toxin from Enteroaggregative Escherichia coli

    doi: 10.1128/IAI.00167-12

    Figure Lengend Snippet: Pet interacts with specific periplasmic proteins from E. coli HB101 during secretion. (A) Bacterial strains grown overnight at 37°C at 150 rpm in 50 ml of LB broth were fractionated as described in Materials and Methods. Cytoplasmic, IM, periplasmic, and OM fractions were analyzed by Western blot (WB) assay using as markers antibodies against GroEL, β-lactamase, and OmpA. (B) The periplasmic fraction of E. coli HB101 was immunoprecipitated with an anti-Pet polyclonal antibody. Experiments were performed with an untransformed laboratory strain that does not express Pet (HB101), a strain transformed with a plasmid encoding wild-type Pet (pCEFN1), a strain transformed with a plasmid encoding the inactive mutant protein PetS260I (pCEFN2), and a strain transformed with a plasmid encoding the truncated mutant Pet protein that lacks the β-barrel translocation unit (pJPN205). An additional negative control involved coimmunoprecipitation of the periplasmic fraction from HB101(pCEFN1) with an isotype IgG control antibody. All immunocomplexes were resolved by SDS-PAGE and visualized by Coomassie blue staining. Arrows indicate the proteins (20, 37 50, and 80 kDa) that specifically interacted with PetS260I; asterisks denote the 50- and 80-kDa proteins that also bound to the truncated mutant Pet protein. MWM, MW markers.

    Article Snippet: The quality of the purified periplasmic fractions was tested by Western blot assay using anti-GroEL (kindly donated by Mario Cancino) and anti-β-lactamase (Chemicon, Temecula, CA) antibodies to detect GroEL (a cytoplasmic protein) and β-lactamase (a periplasmic protein), respectively.

    Techniques: Positron Emission Tomography, Western Blot, Immunoprecipitation, Transformation Assay, Plasmid Preparation, Mutagenesis, Translocation Assay, Negative Control, SDS Page, Staining

    A subset of signal sequences can export thioredoxin-1 to the periplasm. Western analysis of cells (DRH223) expressing signal sequence-thioredoxin fusions fractionated into cytoplasmic and periplasmic fractions by spheroplasting. (A) Cells expressing thioredoxin (TrxA) fused to the AppA (pMO16) and TorT (pMO15) signal sequences (AppAss and TorTss, respectively) from the first iteration of the signal sequence screen. (B) Representative samples from the second iteration of signal sequence screen: TrxA fused to the TolB (pDHSS14), ArtI (pDHSS19), and LivJ (pDHSS29) signal sequences (TolBss, ArtIss, and LivJss, respectively). (C) Representative samples from the third iteration of signal sequence screen: TrxA fused to the FlgA (pDHSS17), YraP (pDHSS18), and YraI (pDHSS20) signal sequences (FlgAss, YraPss, and YraIss, respectively). β-Lactamase is included in all cases as a periplasmic control. W, whole-cell extract; C, cytoplasm plus cytoplasmic membrane fraction; P, periplasm fraction.

    Journal: Journal of Bacteriology

    Article Title: Use of Thioredoxin as a Reporter To Identify a Subset of Escherichia coli Signal Sequences That Promote Signal Recognition Particle-Dependent Translocation

    doi: 10.1128/JB.187.9.2983-2991.2005

    Figure Lengend Snippet: A subset of signal sequences can export thioredoxin-1 to the periplasm. Western analysis of cells (DRH223) expressing signal sequence-thioredoxin fusions fractionated into cytoplasmic and periplasmic fractions by spheroplasting. (A) Cells expressing thioredoxin (TrxA) fused to the AppA (pMO16) and TorT (pMO15) signal sequences (AppAss and TorTss, respectively) from the first iteration of the signal sequence screen. (B) Representative samples from the second iteration of signal sequence screen: TrxA fused to the TolB (pDHSS14), ArtI (pDHSS19), and LivJ (pDHSS29) signal sequences (TolBss, ArtIss, and LivJss, respectively). (C) Representative samples from the third iteration of signal sequence screen: TrxA fused to the FlgA (pDHSS17), YraP (pDHSS18), and YraI (pDHSS20) signal sequences (FlgAss, YraPss, and YraIss, respectively). β-Lactamase is included in all cases as a periplasmic control. W, whole-cell extract; C, cytoplasm plus cytoplasmic membrane fraction; P, periplasm fraction.

    Article Snippet: Rabbit anti-thioredoxin-1 and anti-β-lactamase antibodies for probing membranes were obtained from Sigma and 5′-3′, respectively.

    Techniques: Western Blot, Expressing, Sequencing

    HPLC analysis of the turnover of the cephalosporins cephaloridine (A), cephalothin (B), and cefazolin (C) by EstU1 (green) and class C β-lactamase of E. cloacae (red). The chromatogram of cephalosporins is shown as a black line. The retention

    Journal: Applied and Environmental Microbiology

    Article Title: Novel Metagenome-Derived Carboxylesterase That Hydrolyzes ?-Lactam Antibiotics ▿Novel Metagenome-Derived Carboxylesterase That Hydrolyzes ?-Lactam Antibiotics ▿ †

    doi: 10.1128/AEM.05363-11

    Figure Lengend Snippet: HPLC analysis of the turnover of the cephalosporins cephaloridine (A), cephalothin (B), and cefazolin (C) by EstU1 (green) and class C β-lactamase of E. cloacae (red). The chromatogram of cephalosporins is shown as a black line. The retention

    Article Snippet: Antibiotics (ampicillin, penicillin G, cephaloridine, cephalothin, cefazolin, cefuroxime, and cefotaxime) and class C β-lactamase from Enterobacter cloacae were obtained from Sigma-Aldrich (St. Louis, MO).

    Techniques: High Performance Liquid Chromatography

    Enzyme cyclization using alternative isopeptide-forming domains. ( A ) Schematic of SnoopRing and PilinRing cyclization of β-lactamase (BLA). Proteins are shown in cartoon format, with reacting residues shown in green in stick format. ( B ) SDS-PAGE analysis of SpyRing, SnoopRing and PilinRing cyclization. Each purified protein is shown alongside the non-reactive control (DA or KA), after staining with Coomassie.

    Journal: Scientific Reports

    Article Title: SpyRing interrogation: analyzing how enzyme resilience can be achieved with phytase and distinct cyclization chemistries

    doi: 10.1038/srep21151

    Figure Lengend Snippet: Enzyme cyclization using alternative isopeptide-forming domains. ( A ) Schematic of SnoopRing and PilinRing cyclization of β-lactamase (BLA). Proteins are shown in cartoon format, with reacting residues shown in green in stick format. ( B ) SDS-PAGE analysis of SpyRing, SnoopRing and PilinRing cyclization. Each purified protein is shown alongside the non-reactive control (DA or KA), after staining with Coomassie.

    Article Snippet: For β-lactamase constructs, a final concentration of 100 mM dithiothreitol (DTT, Sigma) was added.

    Techniques: SDS Page, Purification, Staining

    β-lactamase-producing oral anaerobic bacteria

    Journal: Journal of Applied Oral Science

    Article Title: Detection of cfxA2, cfxA3, and cfxA6 genes in beta-lactamase producing oral anaerobes

    doi: 10.1590/1678-775720150469

    Figure Lengend Snippet: β-lactamase-producing oral anaerobic bacteria

    Article Snippet: Isolation and identification of β-lactamase producing bacteria The samples were vortexed for 30 s, serially diluted in phosphate buffered saline, and 0.1 mL of 1/10 to 1/10000 dilutions were plated on 5% blood agar plates supplemented with 5 mg/L of haemin (Sigma-Aldrich, Johannesburg, Gauteng, South Africa) and 1 mg/L of menadione (Sigma-Aldrich, Johannesburg, Gauteng, South Africa) for determining the total anaerobic bacterial count.

    Techniques:

    Rubcn -/- DCs exhibit increased phagosome-to-cytosol escape and proteasome-mediated generation of peptides. Bone marrow-derived dendritic cells (DCs) were generated from Rubcn +/+ (black) and Rubcn -/- (red) mice in vitro with FLT3-L for 7 days. ( A-B ) DCs were loaded with 1 µm CCF4, then co-cultured with apoptotic B16-OVA in the absence or presence of β-lactamase (2 mg/ml) for 90 or 180 minutes at 4°C or 37°C. Uncleaved CCF4 was measured by flow cytometry at an emission of 535 nm, and cleaved CCF4 was measured at an emission of 450 nm. Ratios of 450 nm : 535 nm was calculated by dividing the 450 nm MFI by the 535 nm MFI from a single sample. ( C ) DCs were pre-treated with vehicle or chloroquine (CQ) at 1 or 10 µM for 2 hours and then co-cultured with apoptotic B16-OVA cells (5 apoptotic cells: 1 DC). Eighteen hours later, DCs were harvested for flow cytometry analysis of H2-K b -OVA 257-264 expression. ( D ) DCs were pre-treated with vehicle or MG-132 at 1 or 10 µM for 2 hours and then co-cultured in fresh media with apoptotic B16-OVA cells (5 apoptotic cells: 1 DC). Eighteen hours later, DCs were harvested for flow cytometry analysis of H2-K b -OVA 257-264 expression. ( E ) DCs were pre-treated with vehicle or Brefeldin A at 3 µg/ml for 2 hours and then co-cultured in fresh media with apoptotic B16-OVA cells (5 apoptotic cells: 1 DC). Eighteen hours later, DCs were harvested for flow cytometry analysis of H2-K b -OVA 257-264 expression. Data are expressed as mean ± SEM. No less than two independent experiments were performed, with 3-5 replicates per condition. Significance was calculated using 2-way ANOVA (*p

    Journal: bioRxiv

    Article Title: Non-canonical autophagy in dendritic cells restricts cross-presentation and anti-tumor immunity

    doi: 10.1101/789867

    Figure Lengend Snippet: Rubcn -/- DCs exhibit increased phagosome-to-cytosol escape and proteasome-mediated generation of peptides. Bone marrow-derived dendritic cells (DCs) were generated from Rubcn +/+ (black) and Rubcn -/- (red) mice in vitro with FLT3-L for 7 days. ( A-B ) DCs were loaded with 1 µm CCF4, then co-cultured with apoptotic B16-OVA in the absence or presence of β-lactamase (2 mg/ml) for 90 or 180 minutes at 4°C or 37°C. Uncleaved CCF4 was measured by flow cytometry at an emission of 535 nm, and cleaved CCF4 was measured at an emission of 450 nm. Ratios of 450 nm : 535 nm was calculated by dividing the 450 nm MFI by the 535 nm MFI from a single sample. ( C ) DCs were pre-treated with vehicle or chloroquine (CQ) at 1 or 10 µM for 2 hours and then co-cultured with apoptotic B16-OVA cells (5 apoptotic cells: 1 DC). Eighteen hours later, DCs were harvested for flow cytometry analysis of H2-K b -OVA 257-264 expression. ( D ) DCs were pre-treated with vehicle or MG-132 at 1 or 10 µM for 2 hours and then co-cultured in fresh media with apoptotic B16-OVA cells (5 apoptotic cells: 1 DC). Eighteen hours later, DCs were harvested for flow cytometry analysis of H2-K b -OVA 257-264 expression. ( E ) DCs were pre-treated with vehicle or Brefeldin A at 3 µg/ml for 2 hours and then co-cultured in fresh media with apoptotic B16-OVA cells (5 apoptotic cells: 1 DC). Eighteen hours later, DCs were harvested for flow cytometry analysis of H2-K b -OVA 257-264 expression. Data are expressed as mean ± SEM. No less than two independent experiments were performed, with 3-5 replicates per condition. Significance was calculated using 2-way ANOVA (*p

    Article Snippet: Cells were washed once and co-culture apoptotic B16-OVA cells (5 apoptotic cells : 1 DC) in the absence or presence of 2 mg/ml of purified β-lactamase (Sigma-Aldrich, St. Louis, MO USA) in EM buffer with 1mM probenecid (Sigma-Aldrich, St. Louis, MO USA) for 90 or 180 minutes at 4°C or 37°C.

    Techniques: Derivative Assay, Generated, Mouse Assay, In Vitro, Cell Culture, Flow Cytometry, Expressing

    MgrB localizes to the inner membrane in vivo . (A) The amino acid sequence of MgrB. The underlined region is a potential transmembrane domain [36] . The arrows labeled I and II denote potential type I ( [34] ) and type II [35] signal sequence cleavage sites, respectively. (B) Western blot of the total cell lysate, envelope fraction (pellet), or the soluble protein fraction (sup), of an mgrB − strain expressing cytoplasmic YFP and CFP (AML20) and containing either a control plasmid (pEB52) or an mgrB expression plasmid (pAL8). Both plasmids also express beta-lactamase, an enzyme that resides in the periplasm. (C) Phase contrast and GFP fluorescence micrographs of AML67 ( mgrB − ) expressing either GFP (pAL39, top) or a fusion of GFP to the N-terminus of MgrB (pAL38, bottom). Cells were grown in minimal glucose medium with 1 mM MgSO 4 .

    Journal: PLoS Genetics

    Article Title: Feedback Inhibition in the PhoQ/PhoP Signaling System by a Membrane Peptide

    doi: 10.1371/journal.pgen.1000788

    Figure Lengend Snippet: MgrB localizes to the inner membrane in vivo . (A) The amino acid sequence of MgrB. The underlined region is a potential transmembrane domain [36] . The arrows labeled I and II denote potential type I ( [34] ) and type II [35] signal sequence cleavage sites, respectively. (B) Western blot of the total cell lysate, envelope fraction (pellet), or the soluble protein fraction (sup), of an mgrB − strain expressing cytoplasmic YFP and CFP (AML20) and containing either a control plasmid (pEB52) or an mgrB expression plasmid (pAL8). Both plasmids also express beta-lactamase, an enzyme that resides in the periplasm. (C) Phase contrast and GFP fluorescence micrographs of AML67 ( mgrB − ) expressing either GFP (pAL39, top) or a fusion of GFP to the N-terminus of MgrB (pAL38, bottom). Cells were grown in minimal glucose medium with 1 mM MgSO 4 .

    Article Snippet: Beta-lactamase and CFP/YFP were detected with rabbit polyclonal anti-beta-lactamase (Millipore,) and anti-GFP (A.v.

    Techniques: In Vivo, Sequencing, Labeling, Western Blot, Expressing, Plasmid Preparation, Fluorescence

    The inhibitory activity of quercetin against β-lactamase in hydrolyzing benzylpenicillin. β-lactamase used from E. cloacae ; Con = control (no testing agent); Que (50) = Quercetin 50 μg/mL. The graph shows the remaining benzylpenicillin at the same time. Means sharing the same superscript are not significantly different from each other (Scheffe’s test, p

    Journal: BMC Pharmacology & Toxicology

    Article Title: The synergy and mode of action of quercetin plus amoxicillin against amoxicillin-resistant Staphylococcus epidermidis

    doi: 10.1186/s40360-016-0083-8

    Figure Lengend Snippet: The inhibitory activity of quercetin against β-lactamase in hydrolyzing benzylpenicillin. β-lactamase used from E. cloacae ; Con = control (no testing agent); Que (50) = Quercetin 50 μg/mL. The graph shows the remaining benzylpenicillin at the same time. Means sharing the same superscript are not significantly different from each other (Scheffe’s test, p

    Article Snippet: Amoxicillin, penicillin, β-lactamase type IV, dimethyl sulfoxide (DMSO), glutaraldehyde (Grade I, 25 % for EM), osmium tetroxide (4 % for EM), Spurr Low-Viscosity Embedding Kit and nisin (from Lactococcus lactis , 2.5 % balance sodium chloride and denatured milk solids) were obtained from Sigma (Sigma-Aldrich, UK).

    Techniques: Activity Assay